Correctly diagnosing invasive fungal infections (IFI) in immunocompromised patients is a major unmet medical challenge. Invasive fungal infections are most often caused by various species of Aspergillus and Candida, but also several Fusarium species and Zygomycota such as Mucor, among others. Since the anti-fungal susceptibility of different genera and species differs, specific diagnosis is of great clinical importance. We propose to develop methods for the diagnostic identification of the major pathogenic fungi associated with IFI on the Ibis T6000 biosensor platform. The Ibis T6000 is a novel molecular platform that couples broad-range PCR with electrospray ionization mass spectrometry (ESI-MS). One of the fundamental reasons that conventional PCR methods have not succeeded in fungal diagnostics is the inherent variability of fungal DNA sequences, which leads to difficulty in designing primers and probes that encompass the sequences from broad enough groups of pathogenic fungi to be practical. Ibis technology overcomes these limitations by targeting broadly conserved molecular targets on fungal genomes, which allows for amplification of groups of fungi, including multiple species within a genera and natural variants within a species. Our specific aims are 1) to develop and validate a single test to identify the major fungal pathogens associated with IFI, 2) to develop and validate a single protocol for preparation of fungal nucleic acids from blood and BAL specimens, and 3) to demonstrate identification of Aspergillus fumigatus in blood and BAL using the existing invasive aspergillosis Animal Model (IAAM). If successful, we will then evaluate the Ibis fungal diagnostics test with blood and BAL samples from human subjects with proven and probable invasive aspergillosis in collaboration with the Aspergillus Technology Consortium (AsTeC). Ibis Biosciences has recently (January 2009) been acquired as a subsidiary of Abbott Molecular, Inc. If successful, we will commercialize the fungal diagnostic product using the Abbott worldwide commercial operation.